Fig. 1. NanoLuc complementation (NanoBiT) assay using calmodulin and calmodulin-binding regions. (A) Schematics of cytosolic calcium measurements with NanoBiT assay. Light blue, calmodulin; deep blue, calmodulin target region; red dots, Ca²⁺; red triangle, SmBiT; pink, LgBiT. (B) Optimization of NanoBiT construct combinations. All eight combinations of calmodulin, the target motifs of myosin light chain kinase 1 (MYLK1S), Nluc fragments, and 5-HT2AR were expressed in HEK293T cells. 5-HT (1 μM)-induced luciferase activities were measured as described in the Materials and Methods section. CM, calmodulin; MYLK1S, calmodulin-binding region of MYLK1; Veh., vehicle. All experiments were done more than three times.

Mol. Cells 2020;43:909~920 https://doi.org/10.14348/molcells.2020.0144

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